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- nucleic acid (2)
- nucleophilic addition (2)
- nucleosomes (2)
- nucleotide excision-repair (2)
- nucleotide sequence (2)
- nude-mice (2)
- numerical analysis (2)
- numerical hydrodynamics (2)
- nurse bee (2)
- nurses (2)
- o-methoxy-sila-hexocyclium (2)
- object-based classification (2)
- object-based image analysis (2)
- observer (2)
- obstetrics (2)
- occupational medicine/industrial medicine (2)
- oculomotor control (2)
- oestrogen receptor signalling (2)
- office-workers (2)
- oil palm (2)
- older adults (2)
- olfaktorisches Lernen (2)
- oligodendrocyte (2)
- oligorecurrence (2)
- oligothiophenes (2)
- olive (2)
- omega-3 fatty acids (2)
- oncogene-induced senescence (2)
- oncolytic viral therapy (2)
- oncolytic virus therapy (2)
- one-electron oxidation (2)
- online survey (2)
- onset (2)
- open spaces (2)
- opioid peptides (2)
- optic neuritis (2)
- optical (2)
- optical coherence tomography (2)
- optical physics (2)
- optical remote sensing (2)
- optics (2)
- optics and photonics (2)
- optimal control theory (2)
- optimal drug combination (2)
- optogenetic (2)
- oral bioavailability (2)
- oral mucositis (2)
- ordinary culture (2)
- ordinary differential equations (2)
- organ-on-a-chip (2)
- organic electronics (2)
- organic photovoltaic (2)
- organic thin films (2)
- organische Solarzellen (2)
- origin (2)
- orthopedic surgery (2)
- osteoclasts (2)
- osteonecrosis of the jaw (2)
- osteopontin (2)
- osteoradionecrosis (2)
- otitis media (2)
- outer membrane protein (2)
- outer membrane proteins (2)
- outer-membrane proteins (2)
- outpatients (2)
- oxaliplatin (2)
- oxidative addition (2)
- oxygen reduction reaction (2)
- oxygen uptake (2)
- oxytocin (2)
- p-Halbleiter (2)
- p34 (2)
- p38 MAP kinase (2)
- p44 (2)
- p90 ribosomal S6 kinase (2)
- pCa (2)
- pH (2)
- pICln (2)
- paediatric patients (2)
- paediatrics (2)
- pain regulation (2)
- pain sensation (2)
- paleogeography (2)
- palladium (2)
- palliative care (2)
- pandemic (2)
- panel sequencing (2)
- parabiosis (2)
- parallel evolution (2)
- parallel imaging (2)
- parasite biology (2)
- parasitic cell cycles (2)
- parasitoid (2)
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- parkinson's disease (2)
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- parthenogenesis (2)
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- particles (2)
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- pathogen (2)
- pathogenic bacteria (2)
- pathway analysis (2)
- patient education (2)
- patient reported outcome measures (2)
- patient safety (2)
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- patient-reported outcomes (2)
- patient-specific (2)
- pattern (2)
- peak oxygen uptake (2)
- pedagogical content knowledge (2)
- pediatric adrenocortical cancer (2)
- pediatric adrenocortical tumor (2)
- pediatric patients (2)
- peer review (2)
- pelvic examination (2)
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- pembrolizumab (2)
- pemphigus foliaceus (2)
- penicillin allergy (2)
- penicillin hypersensitivity (2)
- pentacoordinate (2)
- people (2)
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- peptide tyrosine tyrosine (PYY) (2)
- peptide vaccination (2)
- peptides (2)
- performance modeling (2)
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- pericyclic reaction (2)
- pericytes (2)
- periodization (2)
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- peripheral nerve (2)
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- peroxisomes (2)
- personalised medicine (2)
- personality traits (2)
- personalized treatment (2)
- perspective taking (2)
- perspectives (2)
- perylene bisimides (2)
- pest control (2)
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- phage (2)
- phagosome maturation (2)
- pharmaceutical analysis (2)
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- pharmacodynamics (2)
- pharmacokinetic (2)
- pharmacology/toxicology (2)
- pharmacovigilance (2)
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- phenprocoumon (2)
- phosphatidylserine (2)
- phosphodiesterase (2)
- phosphoglycolate phosphatase (2)
- phospholipase D (2)
- phosphorus (2)
- photoemission spectroscopy (2)
- photoinduzierter Elektronentransfer (2)
- photolysis (2)
- photon statistics (2)
- photon-counting (2)
- photoreceptor (2)
- photoreceptors (2)
- photoswitchable organic fluorophores (2)
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- physical health (2)
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- physiologically based pharmacokinetic (PBPK) modeling (2)
- phytochemicals (2)
- phytochemistry (2)
- phytohormones (2)
- pi-conjugation (2)
- piRNA (2)
- pig (2)
- pigmentation (2)
- piperacillin/tazobactam (2)
- piperidines (2)
- placebo-controlled trial (2)
- planning (2)
- plant (2)
- plant cuticle (2)
- plant defence (2)
- plant defenses (2)
- plant diversity (2)
- plant immunity (2)
- plant quality (2)
- plant–insect interactions (2)
- plaque (2)
- plasma proteins (2)
- plasminogen (2)
- platelet physiology (2)
- platelet receptors (2)
- platyhelminthes (2)
- plausibility (2)
- pleomorphic adenoma (2)
- pneumolysin (2)
- pneumonia (2)
- point of care testing (2)
- pointing gestures (2)
- polarimetry (2)
- polaritons (2)
- polarized epithelium (2)
- policy (2)
- politics of representation (2)
- pollen analysis (2)
- pollen metabarcoding (2)
- pollen tube (2)
- pollination network (2)
- pollinator decline (2)
- pollinator friendly plants (2)
- pollinators (2)
- poly(2- oxazoline) (2)
- polycationic peptides (2)
- polymavirus (2)
- polymer-peptide-conjugate (2)
- polymorphisms (2)
- polymyalgia rheumatica (2)
- polyploidy (2)
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- population structure (2)
- population-based (2)
- populations (2)
- pore formation (2)
- porphyrins (2)
- pose estimation (2)
- positive selection (2)
- positron emission tomography/computed tomography (2)
- post-processing (2)
- post-transcriptional regulation (2)
- posterior parietal cortex (2)
- postmenopausal women (2)
- postnatal development (2)
- postoperative bleeding (2)
- postoperative care (2)
- postoperative nausea and vomiting (2)
- posttranslational modifications (2)
- postural control (2)
- potassium channel (2)
- power training (2)
- pp Collisions (2)
- pp collisions (2)
- pre-messenger RNA (2)
- precipitation (2)
- precision agriculture (2)
- preconditioning (2)
- predation risk (2)
- predictive factors (2)
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- priming (2)
- printability (2)
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- probability (2)
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- programmed cell death (2)
- projection neurons (2)
- promoter methylation (2)
- propensity score matching (2)
- propulsion (2)
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- prostate-cancer (2)
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- prosthesis (2)
- protease inhibition (2)
- proteasome (2)
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- psychoeducation (2)
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- puberty (2)
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- pyrene dimer (2)
- pyridoxal phosphatase (2)
- qPCR (2)
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- quantitative SPECT/CT (2)
- quantitative analysis (2)
- quantity discrimination (2)
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- real time PCR (2)
- real world evidence (2)
- receptor beta (2)
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- receptor tyrosine kinase (2)
- reconstructed human epidermis (2)
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- reflection (2)
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- revision arthroplasty (2)
- rheology (2)
- rheumatology (2)
- ribonucleases (2)
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- ribozymes (2)
- rice (2)
- ring expansion (2)
- ring-expansion reaction (2)
- risk profiling (2)
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- robotic (2)
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- rodent model (2)
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- rolipram (2)
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- root (2)
- root growth (2)
- roots (2)
- ruthenium bda complexes (2)
- saccharomyces cerevisiae (2)
- safety behavior (2)
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- salicylic acid (2)
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- sar (2)
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- satellite (2)
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- scaffolds (2)
- scalability (2)
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- secondary site infection (2)
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- see (2)
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- semismooth Newton method (2)
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- sentinel-2 (2)
- septic (2)
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- sesame (2)
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- shape fidelity (2)
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- shoulder (2)
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- similarity (2)
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- sirolimus (2)
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- situation awareness (2)
- sizing (2)
- skeletal dysplasia (2)
- skin model (2)
- skinned fiber (2)
- sleep bruxism (2)
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- smart speaker (2)
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- snow cover duration (2)
- social (2)
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- social context (2)
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- social robot (2)
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- sodium channel (2)
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- sofosbuvir (2)
- solar cell (2)
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- solid-state NMR (2)
- solid-state emitters (2)
- solitary bee (2)
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- soluble guanylyl cyclase (2)
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- somatic mosaicism (2)
- somatostatin receptor (SSTR) (2)
- soybean (2)
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- spatial attention (2)
- spatial navigation (2)
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- spatial presence (2)
- spatial scale (2)
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- speckle tracking (2)
- spectral karyotyping (2)
- speech (2)
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- speed (2)
- sperm DNA methylation (2)
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- spermatocytes (2)
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- spheroids (2)
- sphingosine (2)
- sphingosine kinase inhibitor SKI-II (2)
- sphingosine-1-phosphate (2)
- spider (2)
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- spinal cord injury (2)
- spinal-cord (2)
- spinal-cord-injury (2)
- spine (2)
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- spin–orbit coupling (2)
- spiral ganglion neuron (2)
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- splice variant (2)
- split-belt treadmill (2)
- spontaneous blinks (2)
- spontaneous symmetry breaking (2)
- sporidia (2)
- squaraine dyes (2)
- ssVEP (2)
- stability analysis (2)
- stage renal-disease (2)
- startle (2)
- state (2)
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- stereochemistry (2)
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- structure-activity (2)
- structure-function relationships (2)
- structured illumination (2)
- structured illumination microscopy (2)
- student training (2)
- study design (2)
- sturgeon (2)
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- sub-Saharan Africa (2)
- subcutaneous animal model (2)
- substantia nigra pars compacta (2)
- succession (2)
- suffering (2)
- suicidality (2)
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- sulfates (2)
- sun exposure (2)
- super-resolution fluorescence microscopy (2)
- superparamagnetism (2)
- supervisors (2)
- support vector machines (2)
- supraglacial lakes (2)
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- supramolekulare Chemie (2)
- supraparticles (2)
- surface states (2)
- surfaces, interfaces and thin films (2)
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- survival analysis (2)
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- swimming (2)
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- synapse formation (2)
- synapsin (2)
- synapsis (2)
- synaptische Proteine (2)
- synaptonemal complex (2)
- syndrome (2)
- synergistic effect (2)
- synovial fluid (2)
- synthetic aperture RADAR (2)
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- synthetic mesh (2)
- systematics (2)
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- time (2)
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- tissue model (2)
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- tobacco (2)
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- tolerability (2)
- toll-like receptors (2)
- tool use (2)
- tooth extraction (2)
- top quark decay (2)
- topic modeling (2)
- topologische Isolatoren (2)
- total joint arthroplasty (2)
- toxicology (2)
- toxin (2)
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- tracer (2)
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- tracking (2)
- training intensity distribution (2)
- trait emotional intelligence (2)
- trans-Golgi network (2)
- trans-formanilide (2)
- transcranial magnetic stimulation (2)
- transcription factor MIZ-1 (2)
- transcriptional profiling (2)
- transcriptomic analysis (2)
- transcutaneous vagus nerve stimulation (2)
- transfer learning (2)
- transformation (2)
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- transforming growth factor-beta 1 (2)
- transient dynamics (2)
- transient middle cerebral artery occlusion (2)
- transiente Absorption (2)
- transiente Absorptionsspektroskopie (2)
- transition (2)
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- translation initiation (2)
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- triplet (2)
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- two-dimensional topological insulator (2)
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- two-stage exchange (2)
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- type VII secretion system (2)
- tyramine (2)
- tyrosine (2)
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- ultrafast spectroscopy (2)
- ultrafiltration (2)
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- umbilical hernia (2)
- unconventional superconductivity (2)
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- unfolded protein response (2)
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- unmanned aerial vehicles (2)
- unpleasant pictures (2)
- up-regulation (2)
- update (2)
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- usability (2)
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- vacancy (2)
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- vacuole (2)
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- variant surface glycoprotein (2)
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- vasculature (2)
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- vasopressin (2)
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- velocity-dependent collision frequency (2)
- vemurafenib (2)
- venous system (2)
- ventral hernia (2)
- verdünnt magnetische Halbleiter (2)
- vertebral metastases (2)
- vertebrate (2)
- vertigo (2)
- very-long-baseline interferometry (2)
- vesicle trafficking (2)
- vibration (2)
- vicinal surfaces (2)
- video analysis (2)
- virology (2)
- virtual agent (2)
- virtual body ownership (2)
- virtuelle Realität (2)
- virus reactivation (2)
- viscosity (2)
- visible cross section (2)
- visual (2)
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- vitamin D deficiency (2)
- vitamins (2)
- vocabulary (2)
- volatile anesthetics (2)
- volatiles (2)
- voltage clamp (2)
- volume regulation (2)
- voluntary movement (2)
- voxel-based morphometry (2)
- vulnerability (2)
- wR factor = 0.068 (2)
- wasps (2)
- water balance (2)
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Fluorescence microscopy is a form of light microscopy that has developed during the 20th century and is nowadays a standard tool in Molecular and Cell biology for studying the structure and function of biological molecules. High-resolution fluorescence microscopy techniques, such as dSTORM (direct Stochastic Optical Reconstruction Microscopy) allow the visualization of cellular structures at the nanometre scale (10−9 m). This has already made it possible to decipher the composition and function of various biopolymers, such as proteins, lipids and nucleic acids, up to the three-dimensional (3D) structure of entire organelles. In practice, however, it has been shown that these imaging methods and their further developments still face great challenges in order to achieve an effective resolution below ∼ 10 nm. This is mainly due to the nature of labelling biomolecules. For the detection of molecular structures, immunostaining is often performed as a standard method. Antibodies to which fluorescent molecules are coupled, recognize and bind specifcally and with high affnity to the molecular section of the target structure, also called epitope or antigen. The fluorescent molecules serve as reporter molecules which are imaged with the use of a fluorescence microscope. However, the size of these labels with a length of about 10-15 nm in the case of immunoglobulin G (IgG) antibodies, cause a detection of the fluorescent molecules shifted to the real position of the studied antigen. In dense regions where epitopes are located close to each other, steric hindrance between antibodies can also occur and leads to an insuffcient label density. Together with the shifted detection of fluorescent molecules, these factors can limit the achievable resolution of a microscopy technique. Expansion microscopy (ExM) is a recently developed technique that achieves a resolution improvement by physical expansion of an investigated object. Therefore, biological samples such as cultured cells, tissue sections, whole organs or isolated organelles are chemically anchored into a swellable polymer. By absorbing water, this so-called superabsorber increases its own volume and pulls the covalently bound biomolecules isotropically apart. Routinely, this method achieves a magnifcation of the sample by about four times its volume. But protocol variants have already been developed that result in higher expansion factors of up to 50-fold. Since the ExM technique includes in the frst instance only the sample treatment for anchoring and magnifcation of the sample, it can be combined with various standard methods of fluorescence microscopy. In theory, the resolution of the used imaging technique improves linearly with the expansion factor of the ExM treated sample. However, an insuffcient label density and the size of the antibodies can here again impair the effective achievable resolution. The combination of ExM with high-resolution fluorescence microscopy methods represents a promising strategy to increase the resolution of light microscopy. In this thesis, I will present several ExM variants I developed which show the combination of ExM with confocal microscopy, SIM (Structured Illumination Microscopy), STED (STimulated Emission Depletion) and dSTORM. I optimized existing ExM protocols and developed different expansion strategies, which allow the combination with the respective imaging technique. Thereby, I gained new structural insights of isolated centrioles from the green algae Chlamydomonas reinhardtii by combining ExM with STED and confocal microscopy. In another project, I combined 3D-SIM imaging with ExM and investigated the molecular structure of the so-called synaptonemal complex. This structure is formed during meiosis in eukaryotic cells and contributes to the exchange of genetic material between homologous chromosomes. Especially in combination with dSTORM, the ExM method showed its high potential to overcome the limitations of modern fluorescence microscopy techniques. In this project, I expanded microtubules in mammalian cells, a polymer of the cytoskeleton as well as isolated centrioles from C. reinhardtii. By labelling after expansion of the samples, I was able to signifcantly reduce the linkage error of the label and achieve an improved label density. In future, these advantages together with the single molecule sensitivity and high resolution obtained by the dSTORM method could pave the way for achieving molecular resolution in fluorescence microscopy
Results are presented of cloning cDNA of procine growth hormone, analysis of its primary structure, and creation of a construction capable of expression of this cDNA in Esqheriahia coti cells. It is shown that in the population of mRNA coding porcine growth hormone, heterogeneity is noted which is manifested not only at the level of the nucleotide sequence, but also is reflected in the amino acid sequence of the mature hormone.
Microstructural changes and the understanding of their effect on photocurrent generation are key aspects for improving the efficiency of organic photovoltaic devices. We analyze the impact of a systematically increased amount of the solvent additive diiodooctane (DIO) on the morphology of PBDTTT-C:PC71BM blends and related changes in free carrier formation and recombination by combining surface imaging, photophysical and charge extraction techniques. We identify agglomerates visible in AFM images of the 0% DIO blend as PC71BM domains embedded in an intermixed matrix phase. With the addition of DIO, a decrease in the size of fullerene domains along with a demixing of the matrix phase appears for 0.6% and 1% DIO. Surprisingly, transient absorption spectroscopy reveals an efficient photogeneration already for the smallest amount of DIO, although the largest efficiency is found for 3% DIO. It is ascribed to a fine-tuning of the blend morphology in terms of the formation of interpenetrating donor and acceptor phases minimizing geminate and nongeminate recombination as indicated by charge extraction experiments. An increase in the DIO content to 10% adversely affects the photovoltaic performance, most probably due to an inefficient free carrier formation and trapping in a less interconnected donor-acceptor network.
Organic solar cells have great potential to become a low-cost and clean alternative to conventional photovoltaic technologies based on the inorganic bulk material silicon. As a highly promising concept in the field of organic photovoltaics, bulk heterojunction (BHJ) solar cells consist of a mixture of an electron donating and an electron withdrawing component. Their degree of intermixing crucially affects the generation of photocurrent. In this work, the effect of an altered blend morphology on polaron pair dissociation, charge carrier transport, and nongeminate recombination is analyzed by the charge extraction techniques time delayed collection field (TDCF) and open circuit corrected transient charge extraction (OTRACE). Different comparative studies cover a broad range of material systems, including polymer and small-molecule donors in combination with different fullerene acceptors. The field dependence of polaron pair dissociation is analyzed in blends based on the polymer pBTTT-C16, allowing a systematic tuning of the blend morphology by varying the acceptor type and fraction. The effect of both excess photon energy and intercalated phases are minor compared to the influence of excess fullerene, which reduces the field dependence of photogeneration. The study demonstrates that the presence of neat fullerene domains is the major driving force for efficient polaron pair dissociation that is linked to the delocalization of charge carriers. Furthermore, the influence of the processing additive diiodooctane (DIO) is analyzed using the photovoltaic blends PBDTTT-C:PC71BM and PTB7:PC71BM. The study reveals amulti-tiered alteration of the blend morphology of PBDTTT-C based blends upon a systematic increase of the amount of DIO. Domains on the hundred nanometers length scale in the DIO-free blend are identified as neat fullerene agglomerates embedded in an intermixed matrix. With the addition of the additive, 0.6% and 1% DIO already substantially reduces the size of these domains until reaching the optimum 3% DIO mixture, where a 7.1% power conversion efficiency is obtained. It is brought into connection with the formation of interpenetrating polymer and fullerene phases. Similar to PBDTTT-C, the morphology of DIO-free PTB7:PC71BM blends is characterized by large fullerene domains being decreased in size upon the addition of 3% DIO. OTRACE measurements reveal a reduced Langevin-type, super-second order recombination in both blends. It is demonstrated that the deviation from bimolecular recombination kinetics cannot be fully attributed to the carrier density dependence of the mobility but is rather related to trapping in segregated PC71BM domains. Finally, with regard to small-molecule donors, a higher yield of photogeneration and balanced transport properties are identified as the dominant factors enhancing the efficiency of vacuum deposited MD376:C60 relative to its solution processed counterpart MD376:PC61BM. The finding is explained by a higher degree of dimerization of the merocyanine dye MD376 and a stronger donor-acceptor interaction at the interface in the case of the vacuum deposited blend.
Government funding of research beyond biomedicine: challenges and opportunities for neuroethology
(2022)
Curiosity-driven research is fundamental for neuroethology and depends crucially on governmental funding. Here, we highlight similarities and differences in funding of curiosity-driven research across countries by comparing two major funding agencies—the National Science Foundation (NSF) in the United States and the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG). We interviewed representatives from each of the two agencies, focusing on general funding trends, levels of young investigator support, career-life balance, and international collaborations. While our analysis revealed a negative trend in NSF funding of biological research, including curiosity-driven research, German researchers in these areas have benefited from a robust positive trend in DFG funding. The main reason for the decrease in curiosity-driven research in the US is that the NSF has only partially been able to compensate for the funding gap resulting from the National Institutes of Health restricting their support to biomedical research using select model organisms. Notwithstanding some differences in funding programs, particularly those relevant for scientists in the postdoctoral phase, both the NSF and DFG clearly support curiosity-driven research.
Aging is a one-way process associated with profound structural and functional changes in the organism. Indeed, the neuromuscular system undergoes a wide remodeling, which involves muscles, fascia, and the central and peripheral nervous systems. As a result, intrinsic features of tissues, as well as their functional and structural coupling, are affected and a decline in overall physical performance occurs. Evidence from the scientific literature demonstrates that senescence is associated with increased stiffness and reduced elasticity of fascia, as well as loss of skeletal muscle mass, strength, and regenerative potential. The interaction between muscular and fascial structures is also weakened. As for the nervous system, aging leads to motor cortex atrophy, reduced motor cortical excitability, and plasticity, thus leading to accumulation of denervated muscle fibers. As a result, the magnitude of force generated by the neuromuscular apparatus, its transmission along the myofascial chain, joint mobility, and movement coordination are impaired. In this review, we summarize the evidence about the deleterious effect of aging on skeletal muscle, fascial tissue, and the nervous system. In particular, we address the structural and functional changes occurring within and between these tissues and discuss the effect of inflammation in aging. From the clinical perspective, this article outlines promising approaches for analyzing the composition and the viscoelastic properties of skeletal muscle, such as ultrasonography and elastography, which could be applied for a better understanding of musculoskeletal modifications occurring with aging. Moreover, we describe the use of tissue manipulation techniques, such as massage, traction, mobilization as well as acupuncture, dry needling, and nerve block, to enhance fascial repair.
Microcin C (McC) is a peptide-nucleotide antibiotic produced by Escherichia coli cells harboring a plasmid-borne operon mccABCDE. The heptapeptide MccA is converted into McC by adenylation catalyzed by the MccB enzyme. Since MccA is a substrate for MccB, a mechanism that regulates the MccA/MccB ratio likely exists. Here, we show that transcription from a promoter located upstream of mccA directs the synthesis of two transcripts: a short highly abundant transcript containing the mccA ORF and a longer minor transcript containing mccA and downstream ORFs. The short transcript is generated when RNA polymerase terminates transcription at an intrinsic terminator located in the intergenic region between the mccA and mccB genes. The function of this terminator is strongly attenuated by upstream mcc sequences. Attenuation is relieved and transcription termination is induced when ribosome binds to the mccA ORF. Ribosome binding also makes the mccA RNA exceptionally stable. Together, these two effects-ribosome induced transcription termination and stabilization of the message-account for very high abundance of the mccA transcript that is essential for McC production. The general scheme appears to be evolutionary conserved as ribosome-induced transcription termination also occurs in a homologous operon from Helicobacter pylori.
Bacterial mastitis is caused by invasion of the udder, bacterial multiplication and induction of
inflammatory responses in the bovine mammary gland. Disease severity and the cause of disease are
influenced by environmental factors, the cow’s immune response as well as bacterial traits. Escherichia coli (E. coli) is one of the main causes of acute bovine mastitis, but although pathogenic E. coli strains can be classified into different pathotypes, E. coli causing mastitis cannot unambiguously be distinguished from commensal E. coli nor has a common set of virulence factors
been described for mastitis isolates. This project focussed on the characterization of virulence-
associated traits of E. coli mastitis isolates in comprehensive analyses under conditions either
mimicking initial pathogenesis or conditions that E. coli mastitis isolates should encounter while entering the udder. Virulence-associated traits as well as fitness traits of selected bovine mastitis or faecal E. coli strains were identified and analyzed in comparative phenotypic assays. Raw milk whey was introduced to
test bacterial fitness in native mammary secretion known to confer antimicrobial effects.
Accordingly, E. coli isolates from bovine faeces represented a heterogeneous group of which some
isolates showed reduced ability to survive in milk whey whereas others phenotypically resembled
mastitis isolates that represented a homogeneous group in that they showed similar survival and
growth characteristics in milk whey. In contrast, mastitis isolates did not exhibit such a uniform phenotype when challenged with iron shortage, lactose as sole carbon source and lingual
antimicrobial peptide (LAP) as a main defensin of milk. Reduced bacterial fitness could be related to LAP suggesting that bacterial adaptation to an intramammary lifestyle requires resistance to host
defensins present in mammary secretions, at least LAP.
E. coli strain 1303 and ECC-1470 lack particular virulence genes associated to mastitis isolates. To find out whether differences in gene expression may contribute to the ability of E. coli variants to cause mastitis, the transcriptome of E. coli model mastitis isolates 1303 and ECC-1470 were analyzed to
identify candidate genes involved in bacterium-host interaction, fitness or even pathogenicity during bovine mastitis.
DNA microarray analysis was employed to assess the transcriptional response of E. coli 1303 and
ECC-1470 upon cocultivation with MAC-T immortalized bovine mammary gland epithelial cells to
identify candidate genes involved in bacterium-host interaction. Additionally, the cell adhesion and invasion ability of E. coli strain 1303 and ECC-1470 was investigated. The transcriptonal response to the presence of host cells rather suggested competition for nutrients and oxygen between E. coli and MAC-T cells than marked signs of adhesion and invasion. Accordingly, mostly fitness traits that may also contribute to efficient colonization of the E. coli primary habitat, the gut, have been utilized by the mastitis isolates under these conditions. In this study, RNA-Seq was employed to assess the bacterial transcriptional response to milk whey.
According to our transcriptome data, the lack of positively deregulated and also of true virulence-associated determinants in both of the mastitis isolates indicated that E. coli might have adapted by other means to the udder (or at least mammary secretion) as an inflammatory site. We identified traits that promote bacterial growth and survival in milk whey. The ability to utilize citrate promotes fitness and survival of E. coli that are thriving in mammary secretions. According to our results, lactoferrin has only weak impact on E. coli in mammary secretions. At the same time bacterial determinants involved in iron assimilation were negatively regulated, suggesting that, at least during the first hours, iron assimilation is not a challenge to E. coli colonizing the mammary gland. It has been hypothesized that cellular iron stores cause temporary independency to extracellular accessible iron. According to our transcriptome data, this hypothesis was supported and places iron uptake
systems beyond the speculative importance that has been suggested before, at least during early
phases of infection. It has also been shown that the ability to resist extracytoplasmic stress, by oxidative conditions as well as host defensins, is of substantial importance for bacterial survival in mammary secretions.
In summary, the presented thesis addresses important aspects of host-pathogen interaction and
bacterial conversion to hostile conditions during colonization of the mastitis inflammatory site, the mammary gland.
Neuronal representation and processing of chemosensory communication signals in the ant brain
(2008)
Ants heavily rely on olfaction for communication and orientation and ant societies are characterized by caste- and sex-specific division of labor. Olfaction plays a key role in mediating caste-specific behaviours. I investigated whether caste- and sex-specific differences in odor driven behavior are reflected in specific differences and/or adaptations in the ant olfactory system. In particular, I asked the question whether in the carpenter ant, Camponotus floridanus, the olfactory pathway exhibits structural and/or functional adaptations to processing of pheromonal and general odors. To analyze neuroanatomical specializations, the central olfactory pathway in the brain of large (major) workers, small (minor) workers, virgin queens, and males of the carpenter ant C. floridanus was investigated using fluorescent tracing, immunocytochemistry, confocal microscopy and 3D-analyzes. For physiological analyzes of processing of pheromonal and non-pheromonal odors in the first odor processing neuropil , the antennal lobe (AL), calcium imaging of olfactory projection neurons (PNs) was applied. Although different in total glomerular volumes, the numbers of olfactory glomeruli in the ALs were similar across the female worker caste and in virgin queens. Here the AL contains up to ~460 olfactory glomeruli organized in 7 distinct clusters innervated via 7 antennal sensory tracts. The AL is divided into two hemispheres regarding innervations of glomeruli by PNs with axons leaving via a dual output pathway. This pathway consists of the medial (m) and lateral (l) antenno-cerebral tract (ACT) and connects the AL with the higher integration areas in the mushroom bodies (MB) and the lateral horn (LH). M- and l-ACT PNs differ in their target areas in the MB calyx and the LH. Three additional ACTs (mediolateral - ml) project to the lateral protocerebrum only. Males had ~45% fewer glomeruli compared to females and one of the seven sensory tracts was absent. Despite a substantially smaller number of glomeruli, males possess a dual PN output pathway to the MBs. In contrast to females, however, only a small number of glomeruli were innervated by projection neurons of the m-ACT. Whereas all glomeruli in males were densely innervated by serotonergic processes, glomeruli innervated by sensory tract six lacked serotonergic innervations in the female castes. It appears that differences in general glomerular organization are subtle among the female castes, but sex-specific differences in the number, connectivity and neuromodulatory innervations of glomeruli are substantial and likely to promote differences in olfactory behavior. Calcium imaging experiments to monitor pheromonal and non-pheromonal processing in the ant AL revealed that odor responses were reproducible and comparable across individuals. Calcium responses to both odor groups were very sensitive (10-11 dilution), and patterns from both groups were partly overlapping indicating that processing of both odor classes is not spatially segregated within the AL. Intensity response patterns to the pheromone components tested (trail pheromone: nerolic acid; alarm pheromone: n-undecane), in most cases, remained invariant over a wide range of intensities (7-8 log units), whereas patterns in response to general odors (heptanal, octanol) varied across intensities. Durations of calcium responses to stimulation with the trail pheromone component nerolic acid increased with increasing odor concentration indicating that odor quality is maintained by a stable pattern (concentration invariance) and intensity is mainly encoded in the response durations of calcium activities. For n-undecane and both general odors increasing response dynamics were only monitored in very few cases. In summary, this is the first detailed structure-function analyses within the ant’s central olfactory system. The results contribute to a better understanding of important aspects of odor processing and olfactory adaptations in an insect’s central olfactory system. Furthermore, this study serves as an excellent basis for future anatomical and/or physiological experiments.
Accurate quantification of land use/cover change (LULCC) is important for efficient environmental management, especially in regions that are extremely affected by climate variability and continuous population growth such as West Africa. In this context, accurate LULC classification and statistically sound change area estimates are essential for a better understanding of LULCC processes. This study aimed at comparing mono-temporal and multi-temporal LULC classifications as well as their combination with ancillary data and to determine LULCC across the heterogeneous landscape of southwest Burkina Faso using accurate classification results. Landsat data (1999, 2006 and 2011) and ancillary data served as input features for the random forest classifier algorithm. Five LULC classes were identified: woodland, mixed vegetation, bare surface, water and agricultural area. A reference database was established using different sources including high-resolution images, aerial photo and field data. LULCC and LULC classification accuracies, area and area uncertainty were computed based on the method of adjusted error matrices. The results revealed that multi-temporal classification significantly outperformed those solely based on mono-temporal data in the study area. However, combining mono-temporal imagery and ancillary data for LULC classification had the same accuracy level as multi-temporal classification which is an indication that this combination is an efficient alternative to multi-temporal classification in the study region, where cloud free images are rare. The LULCC map obtained had an overall accuracy of 92%. Natural vegetation loss was estimated to be 17.9% ± 2.5% between 1999 and 2011. The study area experienced an increase in agricultural area and bare surface at the expense of woodland and mixed vegetation, which attests to the ongoing deforestation. These results can serve as means of regional and global land cover products validation, as they provide a new validated data set with uncertainty estimates in heterogeneous ecosystems prone to classification errors.
Based on the strand‐like coordination polymer (CP) type \(^{1}\)\(_{∞}\)[Ln(BSB)\(_{3}\)(py)\(_{2}\)], [BSB]−=bis‐salicylatoborate anion, mixed Eu/Tb‐containing compounds of the constitution \(^{1}\)\(_{∞}\)[Eu\(_{x}\)Tb\(_{1-x}\)(BSB)\(_{3}\)(py)\(_{2}\)] were synthesised ionothermally for a phase width of (x=0.25–0.75) and characterized regarding structure and optical properties. Previously, known only for other lanthanides, the mixed 1D−Eu/Tb‐CPs show excellent options for statistic replacement of the Ln‐cations during synthesis yielding solid solutions. The products are highly luminescent, with the chromaticity being a direct function of the amount of the respective Ln‐ions. Corresponding to an overall addition of emission intensities, the green Tb\(^{3+}\) emission and the red Eu\(^{3+}\) emission allow for a chromaticity control that also includes yellow emission. Control of the luminescence colour renders them suitable examples of the versatility of statistic replacement of metal ions in coordination chemistry. In addition, crystallization of [EMIm]\(_{2}\)[YCl\(_{5}\)(py)] illuminates possible other products of the ionothermal reactions of [EMIm][BSB] with LnCl\(_{3}\) constituted by components not being part of the main CPs.
Extreme value theory is concerned with the stochastic modeling of rare and extreme events. While fundamental theories of classical stochastics - such as the laws of small numbers or the central limit theorem - are used to investigate the asymptotic behavior of the sum of random variables, extreme value theory focuses on the maximum or minimum of a set of observations. The limit distribution of the normalized sample maximum among a sequence of independent and identically distributed random variables can be characterized by means of so-called max-stable distributions.
This dissertation concerns with different aspects of the theory of max-stable random vectors and stochastic processes. In particular, the concept of 'differentiability in distribution' of a max-stable process is introduced and investigated. Moreover, 'generalized max-linear models' are introduced in order to interpolate a known max-stable random vector by a max-stable process. Further, the connection between extreme value theory and multivariate records is established. In particular, so-called 'complete' and 'simple' records are introduced as well as it is examined their asymptotic behavior.
Despite available diagnostic tests and recent advances, diagnosis of pulmonary invasive aspergillosis (IPA) remains challenging. We performed a longitudinal case-control pilot study to identify host-specific, novel, and immune-relevant molecular candidates indicating IPA in patients post allogeneic stem cell transplantation (alloSCT). Supported by differential gene expression analysis of six relevant in vitro studies, we conducted RNA sequencing of three alloSCT patients categorized as probable IPA cases and their matched controls without Aspergillus infection (66 samples in total). We additionally performed immunoassay analysis for all patient samples to gain a multi-omics perspective. Profiling analysis suggested LGALS2, MMP1, IL-8, and caspase-3 as potential host molecular candidates indicating IPA in investigated alloSCT patients. MMP1, IL-8, and caspase-3 were evaluated further in alloSCT patients for their potential to differentiate possible IPA cases and patients suffering from COVID-19-associated pulmonary aspergillosis (CAPA) and appropriate control patients. Possible IPA cases showed differences in IL-8 and caspase-3 serum levels compared with matched controls. Furthermore, we observed significant differences in IL-8 and caspase-3 levels among CAPA patients compared with control patients. With our conceptual work, we demonstrate the potential value of considering the human immune response during Aspergillus infection to identify immune-relevant molecular candidates indicating IPA in alloSCT patients. These human host candidates together with already established fungal biomarkers might improve the accuracy of IPA diagnostic tools.
Although the field of fungal infections advanced tremendously, diagnosis of invasive pulmonary aspergillosis (IPA) in immunocompromised patients continues to be a challenge. Since IPA is a multifactorial disease, investigation from different aspects may provide new insights, helpful for improving IPA diagnosis. This work aimed to characterize the human immune response to Aspergillus fumigatus in a multilevel manner to identify characteristic molecular candidates and risk factors indicating IPA, which may in the future support already established diagnostic assays. We combined in vitro studies using myeloid cells infected with A. fumigatus and longitudinal case-control studies investigating patients post allogeneic stem cell transplantation (alloSCT) suffering from IPA and their match controls.
Characteristic miRNA and mRNA signatures indicating A. fumigatus-infected monocyte-derived dendritic cells (moDCs) demonstrated the potential to differentiate between A. fumigatus and Escherichia coli infection. Transcriptome and protein profiling of alloSCT patients suffering from IPA and their matched controls revealed a distinctive IPA signature consisting of MMP1 induction and LGAL2 repression in combination with elevated IL-8 and caspase-3 levels. Both, in vitro and case-control studies, suggested cytokines, matrix-metallopeptidases and galectins are important in the immune response to A. fumigatus. Identified IPA characteristic molecular candidates are involved in numerous processes, thus a combination of these in a distinctive signature may increase the specificity. Finally, low monocyte counts, severe GvHD of the gut (grade ≥ 2) and etanercept administration were significantly associated with IPA diagnosis post alloSCT. Etanercept in monocyte-derived macrophages (MDM) infected with A. fumigatus downregulates genes involved in the NF-κB and TNF-α pathway and affects the secretion of CXCL10.
Taken together, identified characteristic molecular signatures and risk factors indicating IPA may in the future in combination with established fungal biomarkers overcome current diagnostic challenges and help to establish tailored antifungal therapy. Therefore, further multicentre studies are encouraged to evaluate reported findings.
Context: Patients with primary adrenal insufficiency (PAI) or congenital adrenal hyperplasia (CAH) are at a high risk of adrenal crisis (AC). Glucocorticoid sensitivity is at least partially genetically determined by polymorphisms of the glucocorticoid receptor (GR).
Objectives: To determine if a number of intercurrent illnesses and AC are associated with the GR gene polymorphism \(Bcl\)I in patients with PAI and CAH.
Design and patients: This prospective, longitudinal study over 37.7 ± 10.1 months included 47 PAI and 25 CAH patients. During the study period, intercurrent illness episodes and AC were documented.
Results: The study period covered 223 patient years in which 21 AC occurred (9.4 AC/100 pat years). There were no significant differences between \(Bcl\)I polymorphisms (CC (n=29), CG (n=34) and GG (n=9)) regarding BMI, hydrocortisone equivalent daily dose and blood pressure. We did not find a difference in the number of intercurrent illnesses/patient year among \(Bcl\)I polymorphisms (CC (1.5±1.4/pat year), CG (1.2±1.2/pat year) and GG (1.6±2.2/pat year)). The occurrence of AC was not significantly different among the homozygous (GG) genotype (32.5 AC/100 pat years), the CC genotype (6.7 AC/100 pat years) and the CG genotype (4.9 AC/100 pat years). Concomitant hypothyroidism was the highest in the GG genotype group (5/9), compared to others (CC (11/29) and CG (11/34)).
Conclusions: Although sample sizes were relatively small and results should be interpreted with caution, this study suggests that the GR gene polymorphism \(Bcl\)I may not be associated with the frequencies of intercurrent illnesses and AC.
Besides external characteristics and reading a piece of DNA (barcode), the DNA weight per nucleus (genome size) via flow cytometry is a key value to detect species and hybrids and determine ploidy. In addition, the DNA weight appears to be related to various properties, such as the size of the cell and the nucleus, the duration of mitosis and meiosis and the generation time. Sometimes it is even possible to distinguish between groups or sections, which can lead to new classification of the genera. The variation in DNA weight is also useful to analyze biodiversity, genome evolution and relationships between related taxa. Moreover, it is important to know how large a genome is before one determines the base sequence of the DNA of a plant. Flow cytometry is also important for understanding fundamental processes in plants such as growth and development and recognizing chimeras. In the literature, DNA weight measurements are usually limited to one genus and often only locally (Siljak et al. 2010; Bai et al. 2012). In this study, however, it was decided to investigate all vascular plants from one country. This can also contribute to the protection of rare plants. This study is the first flora in the world whose weight of DNA per nucleus and peak patterns has been determined. More than 6400 plants, representing more than 2350 (sub)species (more than 90%) have been collected, thanks to the help of almost 100 volunteers of Floristisch Onderzoek Nederland (Floron). Multiple specimens of many species have therefore been measured, preferably from different populations, in some cases more than fifty. For 1370 species, these values were not previously published. Moreover, a good number of the remaining 45% are new for The Netherlands. In principle, each species has a fixed weight of DNA per nucleus. It has also been found that, especially between the genera, there are strong differences in the number of peaks that determine the DNA weight, from one to five peaks. This indicates that in a plant or organ there are sometimes nuclei with multiples of its standard DNA weight (multiple ploidy levels). It is impossible to show graphs of more than 2350 species. Therefore, we have chosen to show the peak pattern in a new way in a short formula. Within most genera there are clear differences in the DNA weights per nucleus between the species, in some other genera the DNA weight is hardly variable. Based on about twenty genera that were previously measured completely in most cases (‘t Hart et al. 2003: Veldkamp and Zonneveld 2011; Soes et al. 2012; Dirkse et al. 2014, 2015; Verloove et al. 2017; Zonneveld [et al.] 2000−2018), it can be noted that even if all species of a genus have the same number of chromosomes, there can still be a difference of up to three times in the weight of the DNA. Therefore, a twice larger DNA weight does not have to indicate four sets of chromosomes. Finally, this research has also found clues to examine further the current taxonomy of a number of species or genera.
Poly(A)-binding proteins (PABPs) regulate mRNA fate by controlling stability and translation through interactions with both the poly(A) tail and eIF4F complex. Many organisms have several paralogs of PABPs and eIF4F complex components and it is likely that different eIF4F/PABP complex combinations regulate distinct sets of mRNAs. Trypanosomes have five eIF4G paralogs, six of eIF4E and two PABPs, PABP1 and PABP2. Under starvation, polysomes dissociate and the majority of mRNAs, most translation initiation factors and PABP2 reversibly localise to starvation stress granules. To understand this more broadly we identified a protein interaction cohort for both T. brucei PABPs by cryo-mill/affinity purification-mass spectrometry. PABP1 very specifically interacts with the previously identified interactors eIF4E4 and eIF4G3 and few others. In contrast PABP2 is promiscuous, with a larger set of interactors including most translation initiation factors and most prominently eIF4G1, with its two partners TbG1-IP and TbG1-IP2. Only RBP23 was specific to PABP1, whilst 14 RNA-binding proteins were exclusively immunoprecipitated with PABP2. Significantly, PABP1 and associated proteins are largely excluded from starvation stress granules, but PABP2 and most interactors translocate to granules on starvation. We suggest that PABP1 regulates a small subpopulation of mainly small-sized mRNAs, as it interacts with a small and distinct set of proteins unable to enter the dominant pathway into starvation stress granules and localises preferentially to a subfraction of small polysomes. By contrast PABP2 likely regulates bulk mRNA translation, as it interacts with a wide range of proteins, enters stress granules and distributes over the full range of polysomes.
The larvae of the cabbage root fly induce serious damage to cultivated crops of the family Brassicaceae. We here report the biochemical characterisation of neuropeptides from the central nervous system and neurohemal organs, as well as regulatory peptides from enteroendocrine midgut cells of the cabbage maggot. By LC-MALDI-TOF/TOF and chemical labelling with 4-sulfophenyl isothiocyanate, 38 peptides could be identified, representing major insect peptide families: allatostatin A, allatostatin C, FMRFamide-like peptides, kinin, CAPA peptides, pyrokinins, sNPF, myosuppressin, corazonin, SIFamide, sulfakinins, tachykinins, NPLP1-peptides, adipokinetic hormone and CCHamide 1. We also report a new peptide (Yamide) which appears to be homolog to an amidated eclosion hormone-associated peptide in several Drosophila species. Immunocytochemical characterisation of the distribution of several classes of peptide-immunoreactive neurons and enteroendocrine cells shows a very similar but not identical peptide distribution to Drosophila. Since peptides regulate many vital physiological and behavioural processes such as moulting or feeding, our data may initiate the pharmacological testing and development of new specific peptide-based protection methods against the cabbage root fly and its larva.